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A recombinant multiepitope protein for hepatitis B diagnosis
Marilen Queiroz de Souza1, Alexsandro Sobreira Galdino, José Carlos dos Santos
1Laboratório de Biologia Molecular, Universidade de Brasília, 70910-900 Brasília, DF, Brazil.
Biomed Research International
|December 3, 2013
Summary
This study developed a new recombinant hepatitis B core multiepitope antigen (rMEHB) for improved hepatitis B virus (HBV) diagnosis. The rMEHB protein demonstrated stability and effectiveness in enzyme-linked immunosorbent assays (ELISA).
Area of Science:
- Hepatitis B diagnostics
- Recombinant protein development
- Immunological assays
Background:
- Hepatitis B virus (HBV) infection causes liver inflammation.
- Hepatitis B core antibodies (anti-HBc), including IgM and IgG classes, are key diagnostic markers.
- Anti-HBc can indicate acute infection or past exposure, sometimes being the sole detectable marker.
Purpose of the Study:
- To develop a recombinant hepatitis B core multiepitope antigen (rMEHB) for hepatitis B diagnosis.
- To assess the stability and utility of rMEHB in diagnostic assays.
Main Methods:
- A synthetic gene encoding rMEHB was designed and cloned into a pET21a vector.
- The recombinant protein was expressed in E. coli and purified using Ni-NTA affinity chromatography.
- Circular dichroism spectroscopy was used to evaluate protein stability.
- An enzyme-linked immunosorbent assay (ELISA) was performed using rMEHB with patient sera.
Main Results:
- An induced protein of approximately 21 kDa was successfully produced and purified.
- Circular dichroism confirmed rMEHB is thermally stable at pH 7.0 and 8.0.
- rMEHB was effectively utilized in ELISA to differentiate between positive and negative sera.
Conclusions:
- The developed rMEHB is a stable and promising antigen for hepatitis B diagnosis.
- This recombinant antigen shows potential for use in sensitive and specific ELISA-based diagnostic tests for HBV infection.
